LOCUS       40924_46518        3507 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pwFRT-GSr, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3507)
  AUTHORS   Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
  TITLE     Glyphosate resistance as a novel select-agent-compliant, 
            non-antibiotic-selectable marker in chromosomal mutagenesis of the 
            essential genes asd and dapB of Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 75 (19), 6062-6075 (2009)
  PUBMED    19648360
REFERENCE   2  (bases 1 to 3507)
  AUTHORS   Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Molecular Biology and Bio-Engineering, 
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, 
            Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 3507)
  AUTHORS   Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2009) Molecular Biology and Bio-Engineering, 
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, 
            Honolulu, HI 96822, USA
REFERENCE   4  (bases 1 to 3507)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3507)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2009"; volume: "75"; issue: "19"; 
            pages: "6062-6075"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-OCT-2008) Molecular Biology and Bio-Engineering, University of 
            Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, Honolulu, HI 96822,
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (07-APR-2009) Molecular Biology and Bio-Engineering, University of 
            Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, Honolulu, HI 96822,
            USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     On Apr 7, 2009 this sequence version replaced FJ384986.1.
FEATURES             Location/Qualifiers
     source          1..3507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    104..125
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        140..170
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    178..194
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     202..218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    285..358
                     /label=wild type Flp recognition target
                     /note="wild type Flp recognition target"
     protein_bind    complement(302..349)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      389..430
                     /label=PCS12
                     /note="PCS12"
                     /regulatory_class="promoter"
     RBS             441..463
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             499..939
                     /codon_start=1
                     /gene="gat"
                     /product="Gat"
                     /label=gat
                     /note="glyphosate acetyl transferase; confers resistance to
                     glyphosate"
                     /protein_id="ACJ70055.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            499..939
                     /gene="gat"
                     /label=gat
     protein_bind    complement(995..1042)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(1109..1125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1599..1703
                     /label=AmpR promoter
     CDS             1704..2561
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2735..3323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
       61 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      121 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      181 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
      241 gcctgcaggt cgactctagg atccccgggt accgagctcg aattagcttc aaaagcgctc
      301 tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca agatcccctg
      361 attccctttg tcaacagcaa tggatatcag ccgttgactt agttggtatt tccggaatat
      421 catgctgggt tccgaataat tttgtttaac tttaagaagg agatatacca tgggatcgag
      481 cttcgaaagg acaagcatat gatcgaggtc aagcccatca acgccgagga cacctacgac
      541 ctgcggcatc gggtcctccg gcccaaccag ccgatcgagg cgtgcatgtt cgaaagcgat
      601 ctcacgcgga gcgccttcca cctgggcggg ttctacgggg gcaagctgat ttcggtcgct
      661 tcgttccacc aggccgagca ctcggagctt caggggaaga agcagtacca gctgcgcggt
      721 gtggccaccc tggagggcta ccgggagcag aaggccgggt cgtcgctggt caagcacgcc
      781 gaagagatcc tgcggaagcg gggggccgac atgatctggt gcaacgcccg gacctcggcc
      841 tcggggtact accggaagct ggggttcagc gagcaggggg aggtcttcga caccccgccc
      901 gtggggcccc acatcctgat gtacaagcgg atcacctaac tcgagctaat tccattgagt
      961 aagtttttaa gcacatcagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
     1021 ggaacttcgg aataggaact tcaagatccc caattcgagc tcggtacccg gggatcctag
     1081 agtcgacctg caggcatgca agcttggcac tggccgtcgt tttacaacgt cgtgactggg
     1141 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1201 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1261 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     1321 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     1381 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     1441 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     1501 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1561 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1621 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1681 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1741 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1801 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1861 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1921 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1981 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2041 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2101 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2161 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2221 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2281 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2341 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2401 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2461 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2521 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2581 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2641 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2881 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3481 agtcagtgag cgaggaagcg gaagagc
//
